高效毛细管电泳法分析血样尿样中巴比妥类药物

Determination of Barbiturates in Human Plasma and Urine by High Performance Capillary Electrophoresis

建立了血、尿样品中巴比妥类药物的高效毛细管电泳分离测定方法，测定条件为：运行缓冲液配比：１００ｍｍｏｌ／Ｌ十二烷基硫酸钠－１００ｍｍｏｌ／Ｌ磷酸二氢钠－甲醇－水＝７０∶１５∶５∶１０（ｐＨ值调至７．５５±０．０５），操作电压１７ｋＶ，检测波长２００和２８５ｎｍ，２０ｍｉｎ内７种巴比妥类药物全部得到分离。考察并选择了体液样品的预处理方法，测定了血浆和尿液中６种巴比妥类药物浓度（苯巴比妥、甲基苯巴比妥、异戊巴比妥、硫喷妥、戊巴比妥和速可眠）。测定血药浓度的线性范围为５．０～３５μｇ／ｍｌ，尿样药物浓度线性范围为１．０～８．０μｇ／ｍｌ，最低检测浓度为１．０μｇ／ｍｌ，方法重现性为ＲＳＤ小于１３％。

A method has been developed for the determination of barbiturates (barbital phenobarbital, methyl phenobarbital, amobarbital. thiopental, pentobarbital and secobarbital) in human plasma and urine by micellar electrokinetic capillary chromatography (MEKC). The Uniform Design was used for the optimization of the experiment conditions. The on-column sample stocking has been developed for the MEKC determination of barbiturates. Human plasma (0. 5ml) spiked with 6 barbiturates and internal standard(barbital) was extracted with 3×3ml ether 3 times, while 2ml of spiked urine was extracted with 2×6ml chloroform. The combined extract were evaporated to dryness in a rotating evaporator at 35℃. The residue were dissolved in a special sample solvent (10m mol/L SDS plus 5 m mol/L phosphate at PH 6. 50) to yield on -column sample stacking effect. MEKC measurements were carried out in a buffer containing 15m mol/L phosphate at pH 7. 55 plus 70m mol/L sodium dodecyl sulphate (SDS) and 5%methanol. at 17kV running voltage applied to a 50cm×50μm i. d. (45. 5cm effective length)capillary. Migration-time-calibrated peak areas (AR/MT) were proportional to the concentration of 7 barbiturates in the range from 5. 0 to 35 μg/ml plasma, and from 1. 0 to 8. 0 μg/ml urine. The recovery ranged from 86. 6% to 117. 9 % for 6 barbiturates with RSD less than 13%and the detection limits were 1. 0 μg/ml. By the established method, low level of phenobarbital in two patient's plasma was successfully determined 22 hours after the injection of 100mg of phenobarbital